马蓝叶吲哚苷提取纯化工艺优化及其抑菌活性评价  

Optimization of Extraction and Purification Process of Indican from Baphicacanthus cusia Leaves and Its Antibacterial Activity Evaluation

作　　者：杨潇然 薛兴颖 王成章[1,2] 周昊[1,2] YANG Xiaoran;XUE Xingying;WANG Chengzhang;ZHOU Hao(Institute of Chemical Industry of Forest Products,CAF,Key Lab.of Biomass Energy and Material,Jiangsu Province,Key Lab.of Chemical Engineering of Forest Products,National Forestry and Grassland Administration,National Engineering Research Center of Low-Carbon Processing and Utilization of Forest Biomass,Nanjing 210042,China;Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources,Nanjing Forestry University,Nanjing 210037,China)

机构地区：[1]中国林业科学研究院林产化学工业研究所、江苏省生物质能源与材料重点实验室、国家林业和草原局林产化学工程重点实验室、林木生物质低碳高效利用国家工程研究中心,江苏南京210042 [2]南京林业大学江苏省林业资源高效加工利用协同创新中心,江苏南京210037

出　　处：《林产化学与工业》2024年第3期78-86,共9页Chemistry and Industry of Forest Products

基　　金：中国林科院中央级公益性科研院所基本科研业务费专项资金(CAFYBB2022XB002)。

摘　　要：采用超声波低温提取结合大孔树脂吸附分离技术从马蓝叶中提取富集吲哚苷,在单因素试验基础上通过响应面法优化超声波提取工艺,从6种大孔树脂中筛选最适类型并确定大孔树脂纯化吲哚苷的吸附/解吸最佳工艺条件,制备高纯度吲哚苷提取物;采用96孔板微量法测定了高纯度吲哚苷对金黄色葡萄球菌和大肠杆菌的抑菌活性。研究结果表明:超声波提取的最佳工艺条件为5 g干燥马蓝叶粉末、超声波功率458 W、提取时间29 min、温度32℃、料液比1∶10(g∶mL)、提取2次,在此条件下马蓝叶吲哚苷提取率为(93.15±0.26)%。X-5型大孔树脂对吲哚苷吸附/解吸效果最好,大孔树脂纯化最优条件为:大孔树脂100 g、提取液上样量500 mL、洗脱液乙醇体积分数为40%、洗脱液体积400 mL,该条件下吲哚苷回收率为(82.97±0.30)%,纯度由(18.32±0.18)%提升至(86.43±0.31)%;吲哚苷粗提物对金黄色葡萄球菌和大肠杆菌的最小抑菌质量浓度(MIC)值均为350 mg/L,经纯化后的高纯度吲哚苷提取物对金黄色葡萄球菌和大肠杆菌的MIC值分别为21.88和43.75 mg/L,显示出比阳性对照硫酸庆大霉素(MIC值分别为43.75和87.50 mg/L)更好的抑菌效果。In this study,indican was extracted and enriched from the leaves of Baphicacanthus cusia(Ness)Kuntze using ultrasonic low-temperature extraction and adsorption-separation technology with macroporous resin.The ultrasonic extraction process was optimized by response surface methodology based on the single-factor experiment.The optimal type was selected from six kinds of macroporous resins and the optimal adsorption-desorption process conditions of macroporous resin purification of indican were determined.Meanwhile,the high-purity indican extract was prepared.The minimum inhibitory concentration(MIC)of high-purity indican for Staphylococcus aureus and Escherichia coli was determined using 96 well plate micro method.The results showed that the optimal conditions for ultrasonic extraction process were as follows:the dried B.cusia leaf powder was 5 g,the ultrasound power was 458 W,the extraction time was 29 min,the temperature was 32℃,the solid-liquid ratio was 1∶10(g∶mL),and extracted 2 times.Under these conditions,the extraction rate of indican from B.cusia leaves was(93.15±0.26)%.The X-5 type of microporous resin had the best adsorption-desorption performance of the indican.The optimal process conditions of purification with macroporous resin were as follows:macroporous resin was 100 g,the sample volume was 500 mL,the ethanol volume fraction in the eluent was 40%,and the elution volume was 400 mL.Under these conditions,the recovery rate of indician was(82.97±0.30)%,and the purity increased from(18.32±0.18)%to(86.43±0.31)%.The MIC values of indican crude extract against S.aureus and E.coli both were 350 mg/L.The purified indician exhibited excellent antibacterial activity.The MIC values of high-purity indican extract against S.aureus and E.coli were 21.88 and 43.75 mg/L,respectively,which presented better antibacterial effect than the positive control gentamycin sulfate(MIC values were 43.75 and 87.50 mg/L).

关 键 词：马蓝 吲哚苷 超声波低温提取 大孔树脂纯化 抑菌活性 

分 类 号：TQ35[化学工程]